Fig. 5. Establishment of latently infected cell lines with a full-length HIV provirus. (A) Genome organization of a molecular clone of HIV encoding GFP and containing a frameshift mutation in env. (B) Schematic representation of protocol for enrichment of latently infected cells after infection of Jurkat cells with HIV-R7/E^–/GFP (see text for details). (C) Clonal cell lines isolated using the procedure described above were analyzed for GFP expression under basal and stimulated conditions (24 h treatment with TNF-α). (D) Western blot analysis of four representative Jurkat clones latently infected with HIV-R7/E^–/GFP. Clones were treated for 24 h with TNF-α (10 ng/ml) and cell lysates were analyzed by western blotting using an antiserum from an HIV-infected individual (provided by the NIH AIDS Research and Reagent Reference Program). A predominant band of 55 kDa corresponds to the Gag precursor protein. The same samples were analyzed using an antiserum specific for α-tubulin to ensure equal loading.